02709nas a2200241   4500008004100000022001400041245011900055210006900174260010300243300001400346490000700360520177700367653003802144653001402182653001902196653001102215100002302226700002202249700002402271700002502295700001902320856012802339       2013                        eng d  a0300-908400aHuman mitochondrial NDUFS3 protein bearing leigh syndrome mutation is more prone to aggregation than its wild-type0 aHuman mitochondrial NDUFS3 protein bearing leigh syndrome mutati  a23 RUE LINOIS, 75724 PARIS, FRANCEbELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIERcDEC  a2392-24030 v953 a<p>NDUFS3 is an integral subunit of the Q module of the mitochondrial respiratory Complex-I. The combined mutation (T145I + R199W) in the subunit is reported to cause optic atrophy and Leigh syndrome accompanied by severe Complex-I deficiency. In the present study, we have cloned and overexpressed the human NDUFS3 subunit and its double mutant in a soluble form in Escherichia coli. The wild-type (w-t) and mutant proteins were purified to homogeneity through a serial two-step chromatographic purification procedure of anion exchange followed by size exclusion chromatography. The integrity and purity of the purified proteins was confirmed by Western blot analysis and MALDI-TOF/TOF. The conformational transitions of the purified subunits were studied through steady state as well as time resolved fluorescence and CD spectroscopy under various denaturing conditions. The mutant protein showed altered polarity around tryptophan residues, changed quenching parameters and also noticeably altered secondary and tertiary structure compared to the w-t protein. Mutant also exhibited a higher tendency than the w-t protein for aggregation which was examined using fluorescent (Thioflavin-T) and spectroscopic (Congo red) dye binding techniques. The pH stability of the w-t and mutant proteins varied at extreme acidic pH and the molten globule like structure of w-t at pH1 was absent in case of the mutant protein. Both the w-t and mutant proteins showed multi-step thermal and Gdn-HCI induced unfolding. Thus, the results provide insight into the alterations of NDUFS3 protein structure caused by the mutations, affecting the overall integrity of the protein and finally leading to disruption of Complex-I assembly. (C) 2013 Elsevier Masson SAS. All rights reserved.</p>10aAggregation Fluorescence lifetime10aComplex-I10aLeigh syndrome10aNDUFS31 aJaokar, Tulika, M.1 aPatil, Deepak, P.1 aShouche, Yogesh, S.1 aGaikwad, Sushama, M.1 aSuresh, C., G.  uhttp://library.ncl.res.in/content/human-mitochondrial-ndufs3-protein-bearing-leigh-syndrome-mutation-more-prone-aggregation01887nas a2200181   4500008004100000245014700041210006900188260000800257300002500265490000600290520116900296100001801465700002201483700002201505700002501527700002401552856012901576       2017                        eng d00aHigh-throughput mass spectrometry analysis revealed a role for glucosamine in potentiating recovery following desiccation stress in chironomus0 aHighthroughput mass spectrometry analysis revealed a role for gl  cJUN  aArticle Number: 36590 v73 aDesiccation tolerance is an essential survival trait, especially in tropical aquatic organisms that are vulnerable to severe challenges posed by hydroperiodicity patterns in their habitats, characterized by dehydration-rehydration cycles. Here, we report a novel role for glucosamine as a desiccation stress-responsive metabolite in the underexplored tropical aquatic midge, Chironomus ramosus. Using high-throughput liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QToF-MS) analysis, biochemical assays and gene expression studies, we confirmed that glucosamine was essential during the recovery phase in C. ramosus larvae. Additionally, we demonstrated that trehalose, a known stress-protectant was crucial during desiccation but did not offer any advantage to the larvae during recovery. Based on our findings, we emphasise on the collaborative interplay of glucosamine and trehalose in conferring overall resilience to desiccation stress and propose the involvement of the trehalose-chitin metabolic interface in insects as one of the stress-management strategies to potentiate recovery post desiccation through recruitment of glucosamine.1 aThorat, Leena1 aOulkar, Dasharath1 aBanerjee, Kaushik1 aGaikwad, Sushama, M.1 aNath, Bimalendu, B.  uhttp://library.ncl.res.in/content/high-throughput-mass-spectrometry-analysis-revealed-role-glucosamine-potentiating-recovery